The efficient development of a novel recombinant adenovirus zoster vaccine perfusion production process

Nie, Jianqi; Sun, Yuanzhang; Feng, Kai; Huang, Lingling; Li, Ye; Bai, Zhonghu

doi:10.1016/j.vaccine.2022.02.024

Cited by 4 publications

(2 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This behavior is different from the trend of changes in MOIs = 3 and 6. This result is in accordance with the finding of Nie et al 43 that assessed three experimental CPPs (VCD, MOI, and virus production pH) by the design of experiment (DoE) method, and the robust set point of MOI = 9 was applied in both SFs and 2 L benchtop bioreactor. According to figures A and B, in MOIs = 12 and 15, cell growth has stopped since the first hours of infection, and the trend is downward.…”

Section: Results and Discussionsupporting

confidence: 91%

“…These results are in disagreement with the report, which indicated that the rAd yield could be increased with higher VCD at the time of infection and the optimum VCC = 1.04 × 10 6 cells/mL, MOI = 9, and pH = 7.17 was confirmed in the shake flask in HEK 293 cells. 43 Other study demonstrated that with VCD up to 6−8 × 10 6 cells/mL at infection time, the virus titer up to 1−6 × 10 10 viral particle/mL was obtained in the perfusion bioreactor manufacturing Ad26 with PER.C6 as the host cells. showed that the cells continued to grow even until 48 hpi, and only after that time, the cell growth started to decline.…”

Section: Effect Of the Initial Cell Density On Rad26mentioning

confidence: 94%

See 1 more Smart Citation

High-Titer Recombinant Adenovirus 26 Vector GMP Manufacturing in HEK 293 Cells with a Stirred Single-Use Bioreactor for COVID-19 Vaccination Purposes

Sedighikamal,

Sattarzadeh,

Karimi Mostofi

et al. 2023

ACS Omega

View full text Add to dashboard Cite

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 virus) pandemic has shown the importance of pursuing various vaccine manufacturing strategies. In the present study, the HEK 293 cells were infected with recombinant adenovirus serotype 26 (rAd26), and the effects of critical process parameters (CPPs) including viable cell density (VCD) at infection time (0.5 × 10 6 , 0.8 × 10 6 , 1.4 × 10 6 , 1.8 × 10 6 , and 2.5 × 10 6 cells/mL), the multiplicity of infection (MOI) = 3, 6, 9, 12, and 15, and two aeration strategies (high-speed agitation with a sparging system and low-speed agitation with an overlay system) were investigated experimentally. The results of small-scale experiments in 2 L shake flasks (SF 2L) demonstrated that the initial VCD and MOI could affect the cell proliferation and viability. The results at these experiments showed that VCD = 1.4 × 10 6 cells/mL and MOI = 9 yielded TCID 50 /mL = 10 8.9 , at 72 h post-infection (hpi), while the virus titer at VCD = 0.5 × 10 6 and 0.8 × 10 6 cells/mL was lower compared to that of VCD = 1.4 × 10 6 cells/mL. Moreover, our findings showed that VCDs > 1.8 × 10 6 cells/m with MOI = 9 did not have a positive effect on TCID 50 /mL and MOI = 3 and 6 were less efficient, whereas MOI > 12 decreased the viability drastically. In the next step, the optimized CPPs in a small scale were exploited in a 200 L single-use bioreactor (SUB), with good manufacturing practice (GMP) conditions, at RPM = 25 with an overlay system, yielding high-titer rAd26 manufacturing, i.e., TCID 50 /mL = 10 8.9 , at 72 hpi.

show abstract

Section: Results and Discussionsupporting

confidence: 91%

Section: Effect Of the Initial Cell Density On Rad26mentioning

confidence: 94%